Faculty Bibliography

OBJECTIVE:To report a novel approach to fertility preservation in adolescents with Turner syndrome mosaicism by repeated controlled ovarian stimulation and oocyte cryopreservation. DESIGN/METHODS:Case report. SETTING/METHODS:Academic reproductive medicine center. PATIENT(S)/METHODS:Fourteen-year-old adolescent diagnosed with Turner syndrome mosaicism. INTERVENTION(S)/METHODS:Two cyles of controlled ovarian stimulation and oocyte cryopreservation within 1 year. MAIN OUTCOME MEASURE(S)/METHODS:Recovery of oocytes after controlled ovarian stimulation and oocyte cryopreservation. RESULT(S)/RESULTS:Eleven oocytes were retrieved, of which eight were mature and three were immature during the first cycle. One year later, four mature and three immature oocytes were retrieved after a treatment cycle with even higher gonadotropin doses. All oocytes were cryopreserved by vitrification. CONCLUSION(S)/CONCLUSIONS:Controlled ovarian stimulation and oocyte cryopreservation may be an option for fertility preservation in selected adolescents with Turner syndrome mosaicism and impending ovarian failure.

As cancer survival rates continue to improve, many young adults and children will face infertility after successful treatment of their malignant diseases. Fertility is now recognized as a critical component of quality-of-life for cancer survivors and fertility preservation is an emerging discipline that addresses the need for improving cancer survivors' options to have children later in life. Fertility preservation by established methods is often possible in adult female and male cancer patients before starting their gonadotoxic treatments. In prepubertal children, options are still experimental and most challenging. Embryos, oocytes, sperm, or gonadal tissue (ovarian and testicular) can be cryopreserved and stored at subzero temperatures until the time when the patients are disease-free and wish to start a family. As fertility preservation choices include both established and experimental techniques, a highly individualized approach is required in the management of those patients looking for fertility preservation options.

Aromatase inhibitors can be utilized to minimize oestrogen exposure in breast cancer patients undergoing gonadotrophin stimulation. This retrospective-prospective study determined whether using a gonadotrophin-releasing hormone agonist (GnRHa) trigger instead of human chorionic gonadotrophin (HCG) would reduce oestrogen exposure and improve cycle outcomes in aromatase inhibitor cycles. Seventy-four breast cancer patients who desired fertility preservation, with normal ovarian reserve and < 45 years of age received letrozole 5mg/day plus recombinant FSH 150-300 IU/day for ovarian stimulation. Subjects either received HCG 5000-10,000 IU (n=47) or leuprolide acetate 1mg (GnRHa, n=27) as trigger. Oestradiol measurements were repeated 4 days after the trigger and subjects were evaluated for ovarian hyperstimulation syndrome (OHSS). In the GnRHa group, oestradiol concentrations dropped significantly after the trigger than the HCG group (P=0.013) and there was a lower incidence of OHSS. GnRHa trigger resulted in a higher number and percentage of mature oocytes and a higher number of cryopreserved embryos or oocytes compared with HCG. GnRHa trigger improves outcomes by increasing the yield of mature oocytes and embryos in aromatase inhibitor cycles and also decreases the post-trigger oestradiol exposure as well as OHSS risks in women with breast cancer.

Although still experimental, cryopreservation and transplantation techniques for ovarian tissue have been well described, and a number of successful human pregnancies have occurred. Ovarian cryopreservation is the only fertility preservation procedure that can be offered to prepubertal children, and when cytotoxic treatment is urgent. There are two main approaches for autotransplantation of human ovarian tissue. In the heterotopic transplantation, cortical fragments can be grafted subcutaneously at various sites whereas in orthotopic transplantation cortical pieces are transplanted into its original location. Both approaches have their own advantages and disadvantages. While natural pregnancy can occur in orthotopic transplantation, heterotopic transplantation may be indicated if the pelvis is not suitable for transplantation due to previous radiation or severe scar formation. Furthermore, tissue monitoring may be easier in the heterotopic site. In this article, we reviewed the indications, limitations, risks and transplantation techniques for ovarian tissue.

OBJECTIVE:To assess the indications, safety, utilization, and success of ovarian tissue freezing and transplantation. DESIGN/METHODS:Prospective longitudinal analysis. SETTING/METHODS:Academic medical centers. PATIENT(S)/METHODS:Fifty-nine women who underwent ovarian tissue cryopreservation with a slow freezing technique between May 1997 and March 2008. A follow-up was conducted 36.8 +/- 3.6 months after the procedure. INTERVENTION(S)/METHODS:Ovarian tissue harvesting and cryopreservation. MAIN OUTCOME MEASURE(S)/METHODS:Indications, safety, and utilization rates. RESULT(S)/RESULTS:The mean age (+/- SE) was 26.7 +/-1.2 years (range 4-44 years). The majority of patients had either hematologic malignancies (45.7%) or breast cancer (22%). Of these, 57.6% underwent hematopoietic stem cell transplantation. No complications occurred and no histologic evidence of cancer was found in the harvested tissue. The median length of storage was 3.5 +/- 0.3 years (0.06-10.5 years). Fifty-six of 59 patients have not yet used their ovarian tissue. The reasons for nonutilization were social/personal, being still under treatment, and death in 54%, 38%, and 8%, respectively. Only three women (5.1%) underwent transplantation, two with the heterotopic (abdominal wall) and one with the orthotopic technique. One woman with a heterotopic transplant conceived spontaneously and delivered. Of the three transplants, one ceased function after 9 months and two are still functioning at up to 7 years follow-up. CONCLUSION(S)/CONCLUSIONS:Ovarian tissue harvesting appears to be safe but the experience with ovarian transplantation is still limited due to low utilization. As a result, the true value of this procedure remains to be determined.

PURPOSE/OBJECTIVE:Germline mutations in BRCA genes are associated with breast and ovarian cancer susceptibility. Because infertility is associated with breast and ovarian cancer risks, we hypothesized that the mutations in the BRCA gene may be associated with low response to fertility treatments. METHODS:We performed ovarian stimulation in 126 women with breast cancer by using letrozole and gonadotropins for the purpose of fertility preservation by embryo or oocyte cryopreservation. As surrogates of ovarian reserve, the oocyte yield and the incidence of low response were compared with ovarian stimulation according to BRCA mutation status. RESULTS:Of the 82 women who met the inclusion criteria, 47 women (57%) had undergone BRCA testing, and 14 had a mutation in BRCA genes, of which two were of clinically undetermined significance. In BRCA mutation-positive patients, low ovarian response rate was significantly higher compared with BRCA mutation-negative patients (33.3 v 3.3%; P = .014) and with BRCA-untested women (2.9%; P = .012). All BRCA mutation-positive low responders had BRCA1 mutations, but low response was not encountered in women who were only BRCA2 mutation positive. Compared with controls, BRCA1 mutation- but not BRCA2 mutation-positive women produced lower numbers of eggs (7.4 [95% CI, 3.1 to 17.7] v 12.4 [95% CI, 10.8 to 14.2]; P = .025) and had as many as 38.3 times the odds ratio of low response (95% CI, 4.1 to 353.4; P = .001). CONCLUSION/CONCLUSIONS:BRCA1 mutations are associated with occult primary ovarian insufficiency. This finding may, at least in part, explain the link between infertility and breast/ovarian cancer risks.

Clinical studies of oocyte cryopreservation have gained momentum within the recent years; however, no guidelines have yet been established for patient selection. This article discusses the controversial aspects of selecting candidates for oocyte cryopreservation research.

Two patients with severely diminished ovarian reserve who were refractory to aggressive ovarian stimulation conceived with oocytes from prematurely developing antral follicles after dehydroepiandrosterone supplementation. The first patient had 11 and 14.5 mm, and the second patient had 13 mm antral follicles on cycle days 2 and 3 respectively. In the first case, no ovarian stimulation was performed, while the second case received one dose of gonadotrophins with a gonadotrophin-releasing hormone antagonist. Following very early human chorionic gonadotrophin (HCG) triggering on cycle day 5, when antral follicles reached 15 and 18.5 mm in the first case, and 19 mm in the second case, IVF intracytoplasmic sperm injection treatment resulted in pregnancies in both cases, which are currently ongoing at 35 and 14 weeks of gestation. The results in these patients show that pregnancy can be achieved in poor responder patients with prematurely developing antral follicles following early HCG triggering based on follicle size rather than cycle day, with no or minimal stimulation. Whether DHEA supplementation had any impact on the success of these cycles remains to be determined.

AIM/OBJECTIVE:To determine whether monitoring follicle stimulating hormone (FSH) levels in over-responding patients whom the gonadotropins were withheld would predict pregnancy outcome. METHODS:A group of 33 female infertility patients aged between 20-40 years who had to be coasted during controlled ovarian hyperstimulation were recruited for this study. The FSH concentrations on human chorionic gonadotropin (HCG) day and on the four preceding days were measured to determine the threshold FSH concentration for follicular growth and conception. Mean and standard deviation were used for presenting parametric data. Student's t test and the Mann Whitney U test were used for statistical analysis. Two-tailed P < 0.05 was taken as significant. Receiver-operating characteristic (ROC) curves were performed to identify the cutoff level for FSH and E2 at HCG day and four preceding days. RESULTS:Of 33 cycles, 16 were stimulated with recombinant FSH and 17 with recombinant FSH and human menopausal gonadotropin (HMG). The basal and mean coasting FSH levels of FSH and FSH + HMG cycles did not reveal any differences. FSH concentrations decreased the day after coasting began in 26 cycles and fell by 30.2 +/- 2.7% (range 2-53). In 7 cycles, we observed 34.5 +/- 7.4% (range 6-56) declines in FSH levels beginning 2 days after coasting. Of the 31 women who received HCG, 21 conceived during in vitro fertilization and embryo transfer (IVF-ET). Gestational sac and fetal heart activity were visualized in 14 patients. ROC analysis releaved a cutoff value of 4.9 in FSH level at HCG day discriminative of conception or nonconception. CONCLUSION/CONCLUSIONS:The results from this study show that monitoring serum FSH levels during coasting may be helpful in predicting the pregnancy outcome.

Breast cancer is the most common neoplasm in women and accounts for 26% (182,460) of all new cancer cases among women. With the use of screening mammography and advancement in other diagnostic modalities, many cases of breast cancer now can be diagnosed and treated at early stages of the disease. Unfortunately, adjuvant chemotherapy regimens commonly used in the treatment of breast cancer may cause premature ovarian failure due to their cytotoxic effects on the germ cells in the ovary. Therefore preservation of fertility in breast cancer survivors at reproductive age has become an important quality of life issue. Fertility preservation is a recently emerged field of reproductive medicine that may help protect the reproductive capability of the cancer survivors and allow them to have children in the future. Embryo freezing is the most established fertility preservation strategy. But conventional ovarian stimulation protocols are contraindicated in breast cancer patients because of the rise of estrogen and its metabolites to supraphysiological levels. Recently developed ovarian stimulation protocols with aromatase inhibitor letrozole and tamoxifen appear to provide a safe stimulation with endogenous estrogen levels comparable with those achieved in the natural cycle. Oocyte freezing can be considered in single women and in those who do not wish donor sperm. Ovarian tissue freezing could also be an option in breast cancer patients who do not wish or have a time for an in vitro fertilization cycle, which requires 10 to 14 days of ovarian stimulation.